• Title/Summary/Keyword: RNA polymerase subunit II

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A Novel Approach to Investigating Protein/Protein Interactions and Their Functions by TAP-Tagged Yeast Strains and its Application to Examine Yeast Transcription Machinery

  • Jung, Jun-Ho;Ahn, Yeh-Jin;Kang, Lin-Woo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.631-638
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    • 2008
  • Tandem affinity purification (TAP) method combined with LC-MS/MS is the most accurate and reliable way to study the interaction of proteins or proteomics in a genome-wide scale. For the first time, we used a TAP-tag as a mutagenic tool to disrupt protein interactions at the specific site. Although lots of commonly used mutational tools exist to study functions of a gene, such as deletional mutations and site-directed mutagenesis, each method has its own demerit. To test the usefulness of a TAP-tag as a mutagenic tool, we applied a TAP-tag to RNA polymerase II, which is the key enzyme of gene expression and is controlled by hundreds of transcription factors even to transcribe a gene. Our experiment is based on the hypothesis that there will be interrupted interactions between Pol II and transcription factors owing to the TAP-tag attached at the C-terminus of each subunit of Pol II, and the abnormality caused by interrupted protein interactions can be observed by measuring a cell-cycle of each yeast strain. From ten different TAP-tagged strains, Rpb7- and Rpb12-TAP-tagged strains show severe defects in growth rate and morphology. Without a heterodimer of Rpb4/Rpb7, only the ten subunits Pol II can conduct transcription normally, and there is no previously known function of Rpb7. The observed defect of the Rpb7-TAP-tagged strain shows that Rpb7 forms a complex with other proteins or compounds and the interruption of the interaction can interfere with the normal cell cycle and morphology of the cell and nucleus. This is a novel attempt to use a TAP-tag as a proteomic tool to study protein interactions.

Seven Unrecorded Indigenous Fungi from Mudeungsan National Park in Korea

  • Minseo Cho;Sun Lul Kwon;Young Mok Heo;Young Min Lee;Hanbyul Lee;Changmu Kim;Byoung Jun Ahn;Jae-Jin Kim
    • Mycobiology
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    • v.50 no.4
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    • pp.203-212
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    • 2022
  • Fungi act as important decomposers in the forest environment. They recycle essential nutrients, promote plant growth through mycorrhizal relationships, and act as food for small animals. Samples of 265 indigenous fungal species were collected from Mudeungsan National Park in 2020. These species were identified based on morphological, molecular, and phylogenetic analyses using the internal transcribed spacer (ITS), nuclear large subunit rRNA (LSU), and RNA polymerase II second largest subunit (rpb2) regions. Subsequently, seven species were identified as unrecorded species in Korea: Cordyceps cicadae, Dentocorticium bicolor, Hymenochaete nanospora, Physisporinus crataegi, Rigidoporus piceicola, Russula raoultii, and Scutellinia crinita. This study reveals their detailed macro- and microscopic morphological characteristics with phylogenetic trees to report them as unrecorded species in Korea.

First Report of Pyrenophora tritici-repentis in Wheat (Triticum aestivum L.) in Korea and in vitro Selection of an Effective Fungicide

  • Min-Hye Jeong;Eu Ddeum Choi;Seol-Hwa Jang;Sang-Min Kim;Sook-Young Park
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.277-286
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    • 2023
  • Tan spot, caused by Pyrenophora tritici-repentis, is a major foliar disease in wheat worldwide. In April 2021, tan spot symptoms were observed in a commercial wheat field in Suncheon, Jeonnam Province, Korea, with over 5% of the wheat leaves exhibiting symptoms. These symptoms included oval-shaped tan necrosis surrounded by a bright halo. The three representative isolates exhibited irregular mycelial growth on V8-potato dextrose agar and produced pseudothecia. Based on the concatenated sequence datasets of four multi-genes, including the internal transcribed spacer, large subunit ribosomal RNA, glyceraldehyde-3-phosphate dehydrogenase, and RNA polymerase II second-largest subunit genes, phylogenetic analysis revealed that these three isolates clustered in the same clade as P. tritici-repentis. Results of pathogenicity test indicated that the initial symptoms appeared 5 days post-inoculation (dpi), with typical tan spot symptoms developing at 7 dpi. The pathogen was successfully re-isolated from the symptomatic tissues, thus fulfilling Koch's postulates. Furthermore, we selected three fungicides that effectively inhibited the mycelial growth of P. tritici-repentis by more than 90% in vitro. To the best of our knowledge, this is the first report of tan spot disease in wheat in Korea.

First Report of Xenoroussoella triseptata Isolated from Soil in Korea

  • Jung-Joo Ryu;Seung-Yeol Lee;In-Kyu Kang;Leonid N. Ten;Hee-Young Jung
    • The Korean Journal of Mycology
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    • v.50 no.3
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    • pp.195-204
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    • 2022
  • A fungal strain, designated KNUF-20-NI009, was isolated from soil collected from Gunsan-si, Jeollabuk-do, Korea. The isolate showed cultural features typical of the genus Xenoroussoella. Colonies cultivated on malt extract agar were olivaceous-brown to pale olivaceous-white at the margins, with undersides of dark olivaceous to olivaceous-brown and a white margin. The conidia, with a size range of 2.7-5.1×1.6-3.3 ㎛ ($\bar{x}=3.6\times2.6{\mu}m$, n=50), were globoid to ellipsoid in shape, hyaline when immature, becoming light brown to golden-brown when mature, and characterized by 1 or 2 guttules. Multi-locus sequence analysis based on a combined dataset of internal transcribed spacer regions (ITS), large subunit rDNA (LSU), small subunit rDNA (SSU), translation elongation factor 1-alpha (TEF1α), and RNA polymerase II largest subunit (RPB2) sequences revealed KNUF-20-NI009 to be a strain of Xenoroussoella triseptata. This is the first report of this species in Korea.

First Report of Phaeosphaeria chengduensis Isolated from Gametis jucunda in Korea

  • Soo-Min Hong;Seong-Keun Lim;Young-Kun Kim;Sang Jae Suh;Leonid N. Ten;Seung-Yeol Lee;Hee-Young Jung
    • The Korean Journal of Mycology
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    • v.52 no.2
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    • pp.125-134
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    • 2024
  • A fungal strain, KNUF-4H-A belonging to the genus Phaeosphaeria was isolated from the citrus flower chafer (Gametis jucunda) in Chungcheongbuk-do, Korea. This strain was further identified as Phaeosphaeria chengduensis through phylogenetic analyses based on a concatenated dataset of DNA sequences of internal transcribed spacer (ITS) regions, as well as the small subunit rDNA (SSU), large subunit rDNA (LSU), RNA polymerase II second largest subunit (RPB2), and translation elongation factor 1-α (TEF1-α) genes. The isolate KNUF-4H-A exhibited typical cultural characteristics of P. chengduensis, producing colonies that were flattened, greenish-grey at the edges, grey-white at the center, and dark brown on the reverse side. Phaeosphaeria species have been reported in various ecosystems, including terrestrial and freshwater environments. The isolation of KNUF-4H-A from the citrus flower chafer provides valuable insights into the habitat diversity of Phaeosphaeria. This is the first record of Phaeosphaeria chengduensis in Korea.

Molecular Miology of the Poliovirus (폴리오바이러스의 분자생물학)

  • 최원상
    • Journal of Life Science
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    • v.7 no.4
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    • pp.392-401
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    • 1997
  • The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.

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First Report of Leaf Spot of Datura metel Caused by Alternaria tenuissima in Korea

  • Aktaruzzaman, Md.;Kim, Joon-Young;Afroz, Tania;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.330-333
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    • 2015
  • In June 2013, we collected leaf spot disease samples of Datura metel from Gangneung, Gangwon Province, Korea. The symptoms observed were small circular to oval dark brown spots with irregular in shape or remained circular with concentric rings. We isolated the pathogen from infected leaves and cultured the fungus on potato dextrose agar. We examined the fungus morphologically and confirmed its pathogenicity according to Koch's postulates. The results of morphological examinations, pathogenicity tests, and the rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4), glycerol-3-phosphate dehydrogenase (G3PDH) and the RNA polymerase II second largest subunit (RPB2) gene sequence revealed that the causal agent was Alternaria tenuissima. To the best of our knowledge, this is the first report of leaf spot of D. metel caused by A. tenuissima in Korea as well as worldwide.

Characterization and Pathogenicity of Alternaria vanuatuensis, a New Record from Allium Plants in Korea and China

  • Li, Mei Jia;Deng, Jian Xin;Paul, Narayan Chandra;Lee, Hyang Burm;Yu, Seung Hun
    • Mycobiology
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    • v.42 no.4
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    • pp.412-415
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    • 2014
  • Alternaria from different Allium plants was characterized by multilocus sequence analysis. Based on sequences of the ${\beta}$-tubulin (BT2b), the Alternaria allergen a1 (Alt a1), and the RNA polymerase II second largest subunit (RPB2) genes and phylogenetic data analysis, isolates were divided into two groups. The two groups were identical to representative isolates of A. porri (EGS48-147) and A. vanuatuensis (EGS45-018). The conidial characteristics and pathogenicity of A. vanuatuensis also well supported the molecular characteristics. This is the first record of A. vanuatuensis E. G. Simmons & C. F. Hill from Korea and China.

Characterization of Paecilomyces variotii and Talaromyces amestolkiae in Korea Based on the Morphological Characteristics and Multigene Phylogenetic Analyses

  • Nguyen, Thi Thuong Thuong;Paul, Narayan Chandra;Lee, Hyang Burm
    • Mycobiology
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    • v.44 no.4
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    • pp.248-259
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    • 2016
  • During fungal diversity surveys of the order Eurotiales in Korea, two fungal strains, EML-DG33-1 and EML-NCP50, were isolated from samples of rat dung and fig tree leaf collected at a garden located in Gwangju in 2014. To complete the National Species List of Korea, it is a prerequisite to verify whether many questionable species, which were previously recorded but not confirmed, indeed present in Korea. Herein, the isolates were confirmed as undescribed species, Paecilomyces variotii and Talaromyces amestolkiae based on the combination of morphological and phylogenetic analyses of multigenes including the rDNA internal transcribed spacer, ${\beta}-tubulin$, and RNA polymerase II subunit 2.

Characterization of Three Species of Sordariomycetes Isolated from Freshwater and Soil Samples in Korea

  • Lee, Seo Hee;Park, Hyo Sun;Nguyen, Thuong T.T.;Lee, Hyang Burm
    • Mycobiology
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    • v.47 no.1
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    • pp.20-30
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    • 2019
  • During a survey of fungal diversity in the class Sordariomycetes, 3 fungal strains, CNUFC-KMHY6-1, CNUFC-MSW24-2-11, and CNUFC-GW2S-4 were isolated from soil and freshwater samples, respectively in Korea. The strains were analyzed both morphologically and phylogenetically on the basis of internal transcribed spacer and RNA polymerase II second largest subunit gene sequences. On the basis of their morphology and phylogeny, CNUFC-KMHY6-1, CNUFC-MSW24-2-11, and CNUFC-GW2S-4 isolates were identified as Arcopilus aureus, Memnoniella echinata, and Stachybotrys sansevieriae, respectively. To the best of our knowledge, Ar. aureus and M. echinata have not been previously recorded in Korea, and this is the first report of S. sansevieriae from freshwater niche.